Preparation of a Purified Vaccinia Virus Freed from Soluble Antigens

Abstract

The reported experiments form part of a major study of the pattern of virus particle-associated and of soluble antigens, related to vaccinia infection. To study the antigenic composition of the vaccinia virus particle a purified virus material freed from soluble antigen was required. Principally, the method of purification described is a combination of the one used by Marquardt et al(7) and fraction-ation in a sucrose density gradient. Electron microscopy indicated that very little particu-late impurity was present in the virus material banded in the gradient and washed by centrifugation. In the purified material, containing about 2 mg of virus particles per ml, no hemagglutinin and precipitinogenic factors were demonstrable. Ultrasonic treatment immediately preceding the diffusion-in-gel tests was found important as it increased the reactivity of the antigenic preparations.